Fluorescence microscopy study on the cytoskeletal displacements during sperm differentiation in the bush-cricket Tylopsis liliifolia (Fabricius) (Orthoptera: Tettigoniidae).

A study by fluorescence microscopy has been carried out on male gametes from testicular follicles, seminal vesicles, spermatophores, and seminal receptacles of the bush-cricket Tylopsis liliifolia, focusing the attention on localization and movements of F-actin and α-tubulin during sperm differentiation, since data in this respect are lacking in the Orthoptera. F-actin and α-tubulin positivity was detected in the testicular follicles, in particular at the bridges connecting spermatids of a same clone and around their nucleus, during the first differentiation stages. During the following differentiation stages in the testes, F-actin was found at one of the spermatid poles and then, during nucleus elongation, at the whole acrosomal region. A peculiar F-actin-positivity was found at the flagellum, more markedly immediately posterior to the nucleus, at the basal body region of the gametes from the testicular follicles and from the other examined districts. Other interesting data from our investigations concerns the α-tubulin displacements during the differentiation stages of the spermatid and a constant absence of α-tubulin-positivity where the centrioles are located. No positivity was also found for both α-tubulin and nuclear markers at the anterior region of the gamete, where the acrosomal wings are localized. Our results, compared with what is so far known in literature for the insects, lead us to assert that microfilaments and microtubules undergo gradual displacements, markedly in the testicular follicles, during the morphogenesis of the male gamete of T. liliifolia aimed to its organization and motility and probably also to its interaction with the female gamete.

Spermatodesm reorganization in the spermatophore and in the spermatheca of the bushcricket Tylopsis liliifolia (Fabricius) (Orthoptera, Tettigoniidae).

Spermatozoa of Tettigoniidae are usually transferred to the female by means of a spermatophore which is also the site of feather-shaped spermatodesm formation. These spermatodesms are then transferred to a spermatheca, composed of a spermathecal duct and of a seminal receptacle, involved in storing spermatozoa. In order to extend the knowledge about sperm transfer and spermatodesms reorganization in the Tettigoniidae, a morpho-structural investigation was carried out on spermatophore and spermatheca of Tylopsis liliifolia and on the reorganization of the gametes from the spermatophore. Our results show that the spermatodesms undergo disorganization in the spermatophore; unlike other Tettigoniidae, however, feather-shaped spermatodesms are never found. The epithelium of the spermatheca consists of two cell types, the cuticle-forming and the gland cells, with secretory features. The gland cells, absent in the distal tract of the seminal receptacle, release their secretion in a "reservoir" where an efferent duct opens. In the distal tract of the spermathecal duct, adjacent epithelial cells show diversified ultrastructural characteristics whose probable role is discussed. A particular feature of T. liliifolia is the genesis of the feather-shaped spermatodesms in the seminal receptacle. This feature and the peculiar organization of the feather-shaped spermatodesm are a possible autapomorphy of T. liliifolia.

Light and electron microscopy study of the spermatheca of Eupholidoptera chabrieri bimucronata (Ramme, 1927) and Uromenus brevicollis trinacriae La Greca 1964 (Orthoptera: Tettigoniidae).

A study by both optical and electron microscopy has been carried out on the spermatheca of Eupholidoptera chabrieri bimucronata and Uromenus brevicollis trinacriae (Orthoptera: Tettigoniidae). In both the examined species, the spermatheca consists of a sac/kidney-shaped seminal receptacle and a more or less tortuous spermathecal duct that opens into the common oviduct. The wall of both the organs consists of a pseudostratified epithelium surmounted by a cuticular intima; the latter is made up of a thicker endocuticle and an epicuticle. The epithelium shows two different cell types, irregularly arranged and with well differentiated functions: cuticle-forming and gland cells. In both the species, the cuticle-forming cells perform other functions, in addition to producing the cuticular intima. The gland cells never come in contact with the cuticular intima, have inside the reservoir a secretion whose appearance can diversify also in contiguous zones of the seminal receptacle. Based on our findings in both the species, the functions of the seminal receptacle would differ from those of the spermathecal duct. In the latter, some areas of the wall of the connecting tract show an activity of lysis, by contiguous epithelial cells, that could play a role in control and selection of spermatozoa. As for the feather-shaped spermatodesms, similar in both the species, freeze-fracture observations have shown that the acrosome of each spermatozoon regularly covers three-quarters of the extension of the acrosome of the following spermatozoon. Finally, the significance of our findings, compared with what is known in literature, is discussed.

Sperm storage by spermatodoses in the spermatheca of Trioza alacris (Flor, 1861) hemiptera, psylloidea, triozidae: a structural and ultrastructural study.

Female insects generally store sperm received during mating in specific organs of their reproductive tract, i.e., the spermathecae, which keep the sperm alive for a long time until fertilization occurs. We investigated spermatheca morphology and ultrastructure in the psylloidean insect Trioza alacris (Flor,1861) in which spheroidal sperm packets that we refer to as 'spermatodoses' are found after mating. The ectoderm-derived epithelium of the sac-shaped spermatheca that has a proximal neck, consists of large secretory and flat cuticle-forming cells. Secretory cells are characterized by a wide extracellular cavity, bordered by microvilli, in which electron-dense secretion accumulates before discharge into the spermathecal lumen. The cuticle-forming cells produce the cuticular intima of the organ and a peculiar specialized apical structure, through which secretion flows into the lumen. At mating, the male transfers bundles of sperm cells embedded in seminal fluid into the spermathecal neck. Sperm cells proceed towards the spermathecal sac lumen, where they are progressively compacted and surrounded with an envelope that also encloses secretions of both male and female origin. We describe the formation of these sperm containing structures and document the contribution of the female secretion to spermatodose or female-determined spermatophore construction. We also discuss the choice of the term 'spermatodose' for T. alacris and suggest it be used to refer to sperm masses constructed in the female reproductive organs, at least when they involve the contribution of female secretion.

Structure of male accessory glands of Bolivarius siculus (fischer) (Orthoptera, Tettigoniidae) and protein analysis of their secretions.

In Tettigoniidae (Orthoptera), male reproductive accessory glands are involved in the construction of a two-part spermatophore; one part, the spermatophylax, is devoid of sperm and considered a nuptial gift. The morphology, ultrastructure, and secretion protein content of the male reproductive accessory glands from Bolivarius siculus were investigated. Two main groups of gland tubules open into the ejaculatory duct: the "first-order" glands, a number of large anterior tubules, and the "second-order" glands, smaller and more numerous tubules positioned posteriorly. Along with a further subdivision of the gland tubules, we here describe for the first time an additional gland group, the intermediate tubules, which open between first and second-order glands. The mesoderm-derived epithelium of all glands is a single layer of microvillated cells, which can be either flattened or cylindric in the proximal or distal region of the same gland. Epithelial cells, very rich in RER and Golgi systems, produce secretions of both electron-dense granules and globules or electron-transparent material, discharged into the gland lumen by apocrine or merocrine mechanisms, respectively. With one exception, a unique electrophoresis protein profile was displayed by each of the gland types, paralleling their unique morphologies. To assess the contribution of different types of accessory glands to the construction of the spermatophore, the protein patterns of the gland secretions were compared with those of the extracts from the two parts of the spermatophore. All samples showed bands distributed in a wide range of molecular weight, including proteins of very low molecular mass. However, one major high molecular weight protein band (>180 kDa) is seen exclusively in extracts from the first-order glands, and corresponds to an important protein component of the spermatophylax.

Mode of transfer of spermatozoa in Orthoptera Tettigoniidae.

A morphological and ultrastructural study was carried out on the spermatophore and spermatodoses of some species of Orthoptera Tettigoniidae. From the results concerning the spermatophore it emerged that this structure has a morphological and ultrastructural organization represented by a dilated ampulla and a peduncle or neck. From the examination of freshly deposited spermatophores and those at various time intervals thereafter, it was seen that these structures other than allowing gamete transfer, represent the site where spermatodesms, organized in the male genital tracts, undergo reorganization to acquire their definitive morphological and structural characteristics as found in the female genital tracts. The spermatodoses, in the same way as the spermatophore, represent capsules containing spermatodesms, which are originated in the spermatheca, their specific morphology seems to diversify according to the species considered. As regards their role, it is hypothesized that these structures represent a long-term conservation mechanism for spermatozoa inside the seminal receptacle.